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OBJECTIVE@#To compare the efficacy of eltrombopag combined with cyclosporine A (CsA) and CsA alone in patients with transfusion-dependent non-severe aplastic anemia (TD-NSAA).@*METHODS@#The clinical data of 76 patients with treatment-naive TD-NSAA in Ningde Municipal Hospital of Ningde Normal University and Affiliated Hospital of Nantong University from December 2017 to June 2021 were retrospectively analyzed. Among them, 45 cases were treated with eltrombopag combined with CsA, and 31 patients with compatible baseline characters were treated with CsA alone. The efficacy of patients between the two groups was compared, and the factors affecting the curative effects were also analyzed.@*RESULTS@#There were significant differences in hematological response (HR) and complete response(CR) rates between the two groups at 3, 6, 12 months, and follow-up endpoint of treatment (P<0.05). With the prolongation of eltrombopag treatment time, the curative effect increased gradually, and the patients achieved more CR and HR rates by the end of the follow-up period. Simultaneously, with the increase in the maximum stable dose of eltrombopag, the HR rate increased gradually. The megakaryocyte count in eltrombopag group was higher than that in control at 6 and 12 months (P<0.05). Compared with the control group, the median time of platelet transfusion independence in eltrombopag group was more shorter (P=0.018), and the median platelets transfusion volume was lower (P=0.009). At 3, 6, 12 months after eltrombopag, the change of platelet in eltrombopag group was higher than that in the control group (P<0.05). Analysis of related factors affecting the efficacy showed that sex, age, iron overload, platelet count before treatment had no effect on the efficacy, and the median maximum stable dosage and the administration period for eltrombopag were related to the curative effect. The patients of eltrombopag group experienced adverse events of varying degrees, but the reactions were mild and mostly tolerated.@*CONCLUSION@#Eltrombopag can effectively improve the hematopoietic response and promote platelet recovery for TD-NSAA patients with relatively more residual hematopoietic cells, and it is safe and well tolerated.
Subject(s)
Humans , Anemia, Aplastic/therapy , Retrospective Studies , Treatment Outcome , Cyclosporine/therapeutic use , Immunosuppression Therapy , Immunosuppressive Agents/therapeutic useABSTRACT
@#Abstract: AIDS combined with Pneumocystis jirovecii pneumonia (PJP) and disseminated infections of Talaromyces marneffei and Cryptococcus neoformans are rare. This paper summarizes and analyzes the diagnosis and treatment of an AIDS patient with multiple fungal infections for reference. A 79-year-old male patient was admitted to the hospital with "stool habit change for more than 20 days". The white blood cell count was 4.57×109/L, the percentage of neutrophils was 81.8%, the absolute count of CD4+ lymphocytes was 6/μL, and the CD4/CD8 ratio was 0.17. HIV antibody positive was confirmed by CDC. The cerebrospinal fluid and alveolar lavage fluid were positive for Cryptococcus neoformans capsular antigen, and Pneumocystis jirovecii was found by the bronchoalveolar lavage fluid stained with hexamine silver. The cerebrospinal fluid culture was positive for Cryptococcus neoformans, and the blood culture was positive for Cryptococcus neoformans and Talaromyces marneffei. CT showed that bronchovascular bundles in both lungs were more thick, patchy and cable-like high-density shadows were seen in both lungs, and the edges were blurred. Nodular and cable-like high-density shadows were seen in the posterior apical segment of the left upper lobe, with clear margins. Infection of both lungs was considered, and secondary pulmonary tuberculosis occurred in the left upper lobe. After admission, the patient was treated with various anti-bacterial and fungal drugs due to recurrent fever, but the effect was not effective. The fever symptoms of the patient could not be significantly improved, and his condition continued to worsen, and he eventually died. The patient with AIDS complicated with bacterial and fungal infection, especially PJP infection in serious condifiton and has a poor prognosis for rapid development, so clinical attention should be paid to.
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Objective@#To observe the effects of oxymatrine(OM) on the expression and release of high mobility group box 1(HMGB1) in rat pancreatic acinar cell line AR42J stimulated by hydrogen peroxide.@*Methods@#MTT method was used to detect the effects of H2O2 in different concentrations on the survival of AR42J cells. AR42J cells cultured in vitro were divided into control group, H2O2 group and H2O2+ OM group. An equal volume of H2O2(final concentration 0.16 mmol/L) was added in H2O2 group and H2O2+ OM group, respectively, while an equal volume of triple distilled water was added in control group. In H2O2+ OM group, OM(final concentration 0.5 g/L)was added 0.5 h before the addition of H2O2, and cell samples and supernatant were collected after 24 h culture. The expression of HMGB1 protein was detected by Western blotting, the level of HMGB1 protein in cell supernatant was detected by enzyme-linked immunosorbent assay, and the intracellular distribution of HMGB1 protein was detected by immunofluorescence.@*Results@#In the H2O2 group, the expression of HMGB1, the secretion of HMGB1 in the supernatant and the proportion of cytoplasmic HMGB1 in the total HMGB1 were significantly higher than those in the control group[1.04±0.04 vs 0.69±0.02, (4.84±0.13)μg/L vs (2.68±0.07)μg/L, (35.7±2.5)% vs (10.7±1.9)%], and all the differences were statistically significant (all P<0.05). After OM intervention, HMGB1 protein expression, secretion and cytoplasmic proportion were [0.82±0.02, (3.97±0.10)μg/L and (27.3±1.7)%], respectively, which were obviously lower than those in the H2O2 group, and all the differences were statistically significant (all P<0.05).@*Conclusions@#H2O2 can induce the expression and release of HMGB1 in rat pancreatic acinar cells; OM treatment could alleviate the severity of oxidative stress injuries induced by H2O2 in AR42J cells.
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Objective:To observe the effects of oxymatrine(OM) on the expression and release of high mobility group box 1(HMGB1) in rat pancreatic acinar cell line AR42J stimulated by hydrogen peroxide.Methods:MTT method was used to detect the effects of H 2O 2 in different concentrations on the survival of AR42J cells. AR42J cells cultured in vitro were divided into control group, H 2O 2 group and H 2O 2+ OM group. An equal volume of H 2O 2(final concentration 0.16 mmol/L) was added in H 2O 2 group and H 2O 2+ OM group, respectively, while an equal volume of triple distilled water was added in control group. In H 2O 2+ OM group, OM(final concentration 0.5 g/L)was added 0.5 h before the addition of H 2O 2, and cell samples and supernatant were collected after 24 h culture. The expression of HMGB1 protein was detected by Western blotting, the level of HMGB1 protein in cell supernatant was detected by enzyme-linked immunosorbent assay, and the intracellular distribution of HMGB1 protein was detected by immunofluorescence. Results:In the H 2O 2 group, the expression of HMGB1, the secretion of HMGB1 in the supernatant and the proportion of cytoplasmic HMGB1 in the total HMGB1 were significantly higher than those in the control group[1.04±0.04 vs 0.69±0.02, (4.84±0.13)μg/L vs (2.68±0.07)μg/L, (35.7±2.5)% vs (10.7±1.9)%], and all the differences were statistically significant (all P<0.05). After OM intervention, HMGB1 protein expression, secretion and cytoplasmic proportion were [0.82±0.02, (3.97±0.10)μg/L and (27.3±1.7)%], respectively, which were obviously lower than those in the H 2O 2 group, and all the differences were statistically significant (all P<0.05). Conclusions:H 2O 2 can induce the expression and release of HMGB1 in rat pancreatic acinar cells; OM treatment could alleviate the severity of oxidative stress injuries induced by H 2O 2 in AR42J cells.
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ObjectiveTo systematically evaluate the clinical effect of epidermal growth factor receptor (EGFR)-targeted agents in the treatment of advanced pancreatic cancer. MethodsEMbase, PubMed, Cochrane Library, Clinical Trials, CNKI, Wanfang Data, and VIP were searched for randomized controlled trials (RCTs) of EGFR-targeted therapies for advanced pancreatic cancer. Eligible RCTs were screened out based on quality and related criteria, and RevMan 5.3 and STATA 12.0 were used to perform the meta-analysis. ResultsA total of 8 RCTs were included, with 2382 patients in total. The results of the meta-analysis showed that compared with the control group, the experimental group had no increases in overall survival time (hazard ratio ［HR］=0.86, 95% confidence interval ［CI］: 0.74-1.02, P＞0.05) and objective response rate (risk ratio ［RR］=1.00, 95%CI: 0.76-1.32, P＞0.05), but had significant increases in progression-free survival time (HR=0.78, 95%CI: 0.62-0.98, P＜0.05) and disease control rate (RR=1.22, 95%CI: 1.04-1.43, P＜0.05). The subgroup analysis showed that after the source of heterogeneity was identified and the studies with high heterogeneity were excluded, the experimental group had a significant increase in overall survival time (HR=0.85, 95%CI: 0.77-0.94, P＜0.05), and the patients with rash appeared to be more sensitive to the therapies (HR=0.70, 95%CI: 0.54-0.92, P＜0.05). ConclusionFor patients with advanced pancreatic cancer, EGFR-targeted therapies can effectively prolong overall survival time and improve quality of life.
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Wu-tou decoction (WTD) was originally recorded in the synopsis of the golden chamber and it had been widely used for the treatment of neuropathic pain (NP) with exact therapeutic efficacy. However, the underlying molecular mechanisms still remain unclarified. Thus, in this research, we aimed at clarifying the underlying molecular mechanisms of WTD against NP by combining network analysis and experimental validation based on the spinal nerve ligation (SNL) model. Firstly, the network analysis indicated that key targets of WTD were significantly involved in the MAPK signaling pathway (P = 4.04E-12) and four important components of the above pathway, AKT kinase (AKT), MAP kinase kinase 4 (MKK4), c-Jun N-terminal kinase (JNK) and transcription factor AP-1 (JUN) had been reported to play a vital role in neuroinflammation during the disease process of NP. Then, experimental validation results proved that WTD markedly reduce the severity of mechanical allodynia (P<0.01) and cold hypersensitivity (P<0.05) of SNL rats. In addition, Western blot results provided evidence that the phosphorylated protein expression levels of AKT, MKK4, JNK and JUN in the superficial lamina of spinal cord of SNL rats were markedly increased (P<0.001), and WTD could improve the phosphorylated protein expression level of AKT (P<0.001) which was reported to be nerve protective and attenuate the phosphorylated protein expression levels of MKK4, JNK and JUN (P<0.01) which were closely involved into neuroinflammation. In conclusion, this study indicated that WTD might exert anti-hyperalgesia action through the inhibition of neuroinflammation mediated by AKT-MKK4-JNK-JUN which belong to the MAPK signaling pathway. These findings also provided scientific evidences that WTD might be a promising candidate for NP. Animal experiments in this study were approved by the Ethics Committee of Experimental Animals of the China Academy of Chinese Medical Sciences.
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ObjectiveTo investigate the expression and clinical value of ZNF580 mRNA in hepatocellular carcinoma (HCC), as well as the role of ZNF580 mRNA in the development and progression of HCC. MethodsHCC dataset was downloaded from The Cancer Genome Atlas (TCGA) to obtain the gene expression profile and clinical information of ZNF580 mRNA. The bioinformatics method was used to analyze the association between the expression of ZNF580 mRNA in HCC and clinicopathological indices and the influence of ZNF580 mRNA on prognosis. A gene enrichment analysis was used to predict the possible pathways of ZNF580 mRNA involved in the regulation of HCC. The t-test was used for comparison of continuous data between two groups, the chi-square test was used for comparison of categorical data between two groups. The Cox proportional-hazards regression model was used to investigate the risk factors for prognosis, and hazard ratio (HR) and 95% confidence interval (CI) were calculated. ResultsIn the TCGA dataset, the expression of ZNF580 mRNA in tumor tissue was significantly higher than that in adjacent tissue (8.440±0.705 vs 7.736±0.387, P＜0.05), and there was a significant difference in the expression of ZNF580 mRNA between the patients with different tumor grades (t=-2.068, P＜0.05). The patients with high expression of ZNF580 mRNA had a significantly shorter overall survival time than those with low expression (χ2=5.456, P＜0.05). The multivariate Cox regression analysis showed that ZNF580 mRNA expression ［HR(95%CI)=1.600(1.079~2.372)］ and TNM stage ［HR(95%CI)=2.006(1.394~2.888)］ were independent risk factors for overall survival time of HCC patients (both P＜0.05). The samples with high expression of ZNF580 mRNA showed enrichment of genes in the pathways of ribosome and Huntington’s disease (all P＜0.05, FDR=0.198,0.243). ConclusionZNF580 mRNA may act as an oncogene in HCC and is expected to become an indicator for prognostic evaluation of HCC and a potential therapeutic target for HCC.
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In 1980s, Biomphalaria straminea, an intermediate host of Schistosoma mansoni, was found in Shenzhen City, Guangdong Province, China, and currently, this snail has colonized in Shenzhen City and spread to peripheral cities involving of Dongguan and Huizhou. Since imported cases infected with S. mamoni have been reported from time to time in China, Mainland China is facing the potential risk of transmission of schistosomiasis mansoni. With the deepening of the opening-up policy, notably the implementation of the Belt and Road Initiative, there is an increase in the risk of transmission of schistosomiasis mansoni in Mainland China. Increasing the understanding on schistosomiasis mansoni, improving the awareness toward schistosomiasis mansoni prevention and control, and identifying, reporting and managing imported cases with S. mansoni infection or pathogen carriers, are of particular importance to prevent the development of entire life cycle of S. mansoni and the resultant schistosomiasis mansoni transmission in China. To protect public health, a consensus has been reached pertaining to the surveillance and control strategy of imported schistosomiasis mansoni by Chinese infectious disease experts and parasitologists, with aims to improve the awareness and capability for the diagnosis, treatment and control of imported schistosomiasis mansoni among Chinese disease control and prevention institutions and medical institutions, and decrease and even eliminate the risk of schistosomiasis mansoni transmission in China.
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In 1980s, Biomphalaria straminea, an intermediate host of Schistosoma mansoni, was found in Shenzhen City, Guangdong Province, China, and currently, this snail has colonized in Shenzhen City and spread to peripheral cities involving of Dongguan and Huizhou. Since imported cases infected with S. mamoni have been reported from time to time in China, Mainland China is facing the potential risk of transmission of schistosomiasis mansoni. With the deepening of the opening-up policy, notably the implementation of the Belt and Road Initiative, there is an increase in the risk of transmission of schistosomiasis mansoni in Mainland China. Increasing the understanding on schistosomiasis mansoni, improving the awareness toward schistosomiasis mansoni prevention and control, and identifying, reporting and managing imported cases with S. mansoni infection or pathogen carriers, are of particular importance to prevent the development of entire life cycle of S. mansoni and the resultant schistosomiasis mansoni transmission in China. To protect public health, a consensus has been reached pertaining to the surveillance and control strategy of imported schistosomiasis mansoni by Chinese infectious disease experts and parasitologists, with aims to improve the awareness and capability for the diagnosis, treatment and control of imported schistosomiasis mansoni among Chinese disease control and prevention institutions and medical institutions, and decrease and even eliminate the risk of schistosomiasis mansoni transmission in China.
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OBJECTIVE@#To evaluate the clinical efficacy of low dose combined chemotherapy（LDCC） for patients with relapsed and refractory aplastic anemia-paroxysmal nocturnal hemoglobinuria（AA-PNH） syndrome, and to analyze the advantages of LDCC in the treatment of AA-PNH syndrome.@*METHODS@#The clinical characteristics and the curative effect of LDCC in 9 patients with relapsed and refractory AA-PNH syndrome were retrospectively analyzed. Five patients were treated with MP therapy［melphalan 2 mg/(m·d); prednisone 0.5 mg/(kg·d)］, and the other 4 patients were treated with HA therapy(HHT 2 mg/d iv drip, for 5 days; Ara-C 100 mg/d iv drip, for 5 days). The changes of PNH clone, dosage of corticosteroid, hemolysis and the relapse of disease, hematological parameters and adverse reactions were compared before and after therapy. All patients were treated for 1-2 courses.@*RESULTS@#Seven out of 9 patients responded well, the dosage of corticosteroid and the bilirubin concentration decreased significantly and anemia was relieved in 7 patients (P<0.05). One patient relapsed in one year. PNH clone of 3 patients turned negative. Five patients did not rely on blood transfusion in 1 year. There was no bone marrow failure to be found in all patients.@*CONCLUSION@#The LDCC has better efficacy and safety in the treatment of patients with AA-PNH syndrome, moreover, the patients is more tolerant to LDCC, thus the LDCC may be a selection for treatment of patients with relapsed and refractory AA-PNH syndrome.
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Humans , Anemia, Aplastic , Anemia, Refractory , Hemoglobinuria, Paroxysmal , Hemolysis , Retrospective StudiesABSTRACT
Objective:To study the effect ofmetformin combined with insulin aspart on the serum cholesterol(TC),total Bilirubin (TBil),uric Acid(UA),urinary Micro Protein(mAlb) levels and Maternal and Infant Outcomes of gravida with Gestational Diabetes Mellitus.Methods:84 patients ofgestational diabetes mellitus who received therapy from June 2014 to June 2016 in our hospital were selected.According to random number table,those patients were divided into the observation group (n=42) and the control group (n=42),on the basis of routine treatment,The control group was treated with insulin aspart,while the observation group was combined with metformin hydrochloride.The blood glucose index and the levels of TC,TBil,UA,mAlb and maternal and infant outcomes were compared.Results:After treatment,the levels of fasting blood glucose (FBG),postprandial 2h blood glucose (2hPG),glycosylated hemoglobin (HbA1c),TC,TBil,UA and mAlb in the observation group were significantly lower than the control group,the levels of TBil was significantly higher than the control group (P＜0.05);the incidence ofgestational hypertension,hydramnios,premature birth,cesarean section,giant child and neonatal jaundice were significantly lower than the control group (P ＜0.05).Conclusion:Metformin combined with insulin aspart was well for gestational diabetes mellitus,which could effectively improve the blood glucose indicators and TC,TBil,UA,mAlb levels,maternal and infant outcomes.
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Objective To investigate the risk factors for recurrence after surgery for biliary pancreatitis.Methods A total of 284 patients with biliary pancreatitis who were treated in The Fourth Hospital of PLA from January 2008 to December 2014 were followed up,and the risk factors for postoperative recurrence were analyzed.The chi-square test was used for categorical data,and the unconditional logistic regression model was used for multivariate analysis.Results The follow-up period ranged from 6 to 27 months (mean 36.4 ± 8.4 months).Of all patients,27 experienced recurrence,and the recurrence rate was 9.51％.The univariate analysis showed that postoperative recurrence was associated with a family history of gallstone disease,high-fat diet,sand-like stones,intrahepatic bile duct stones,biliopancreatic duct opening stenosis,and diverticulum around the ampullar region (x2 =8.721,5.979,8.641,15.996,33.833,and 27.203,all P ＜0.05).The multivariate logistic analysis showed that high-fat diet (OR =2.296,P =0.012),biliopancreatic duct opening stenosis (OR =2.280,P =0.007),and diverticulum around the ampullar region (OR =2.522,P =0.009) were independent risk factors for recurrence after surgery for biliary pancreatitis.Conclusion Biliary pancreatitis patients with high-fat diet,biliopancreatic duct opening stenosis,or diverticulum around the ampullar region tend to experience recurrence after surgery.Intervention and close follow-up should be performed for these patients to prevent recurrence.
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Objective:This paper explores the development characteristics of posttraumatic growth,and the acceleration of resiliency in posttraumatic growth among adolescents after Wenchuan Earthquake.Methods:We constructed our sample which consisted of 46 adolescents who had traumatic experience in the Wenchuan Earthquake using stratified sampling method.We tracked and monitored the psychological well-being of our subjects for approximately two and half years and six months using questionnaires.9 individuals from the sample were selected based on purposeful sampling according to the longitudinal data and in-depth interviews were conducted for these selected adolescents.The data were collected by semi-structured interviews,organized by Nvivo8.0 and analyzed using thematic analysis.Results:Posttraumatic growth contained four dimensions,including new possibility of the emergence,positive changes of attitude in interpersonal relationship and in philosophy of life,personal strength except altruistic behavior and spiritual change.Protective factors of resiliency among the interviewed students showed individual and gender differences,adolescents with a higher level of resilience had a higher level of posttraumatic growth.Conclusion:The dimensions of posttraumatic growth have unique features.Resilience plays an important role in facilitating individual's posttraumatic growth after the Wenchuan Earthquake.Positive experience in the past and the positive reflection on the event can promote the level of posttraumatic growth.
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Objective To investigate the safety and effectiveness of endovascular embolization combined with two-stage stereotacticradiosurgery (SRS)for the treatment of intracranial arteriovenous malformation (AVM). Methods From January 2010 to December 2012,the clinical data of 66 patients treated with endovascular embolization combined with two-stage SRS AVM comprehensive therapy at the Department of Neurosurgery,Changhai Hospital,the Second Military Medical University were analyzed retro-spectively. They were followed up by imaging. The followed-up time was 7 to 96 months. DSA revealed that the complete disappearance of vascular flow void shadow was regarded as a cure. The patients were divided into either a cured group (n = 29)or a not cured group (n = 37)according whether they were cured or not. The effects of different factors,such as AVM rupture or not,the volume size before embolization,and the degree of embolization on the cure rate of AVM with the comprehensive therapy were observed in both groups,and logistic regression was used to analyze the factors affecting the cure rate of comprehensive therapy. Results (1)There were 22 patients (75. 9%)had ruptured AVMs and 7 (24. 1%)had unruptured AVMs in the cured group. The diameters of the aneurysms < 3 cm and ≥3 cm were in 26 (89. 7%)and 3 (10. 3%)cases respectively. Spetzler-Martin (SM)grades of patients were Ⅰ to Ⅲ. In the not cured group,20 patients (54. 1%)had ruptured AVMs and 17 (45. 9%)had unruptured AVMs. The diameters< 3 cm and ≥3 cm were 18 (48. 6%)and 19 patients (51. 4%)respectively. The SM grade in 34 cases were Ⅰ to Ⅲ. (2)During the follow-up period,29 patients (43. 9%)achieved cure on imaging. The modified Rankin scale (mRS)scores in 64 cases (97%)were 0 to 1. Six patients had complications. (3)Multivariate logistic regression analysis showed that the size of AVM (OR,0. 141,95% CI 0. 035 -0. 570,P < 0. 01)and the degree of interventional embolization (OR,2. 414,95% CI 1. 038 -5. 613,P <0. 05)were the influencing factor of the cure rate. Conclusions Vascular interventional embolization in combination with SRS for the treatment of intracranial AVMs was both effective and safe. The diameter of AVM < 3 cm was the beneficial factor of cure rate of comprehensive therapy. The degree of interventional embolization not reaching cure on imaging was a risk factor for the cure rate of comprehensive therapy.
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Angiostrongyliasis is difficult to be diagnosed for the reason that no ideal method can be used. Serologic tests require specific equipment and are not always available in poverty-stricken zone and are time-consuming. A lateral flow immunoassay (LFIA) may be useful for angiostrongyliasis control. We established a LFIA for the diagnosis of angiostrongyliasis based on 2 monoclonal antibodies (mAbs) against antigens of Angiostrongylus cantonensis adults. The sensitivity and specificity were 91.1% and 100% in LFIA, while those of commercial ELISA kit was 97.8% and 86.3%, respectively. Youden index was 0.91 in LFIA and 0.84 in commercial ELISA kit. LFIA showed detection limit of 1 ng/ml of A. cantonensis ES antigens. This LFIA was simple, rapid, highly sensitive and specific, which opened an alternative approach for the diagnosis of human angiostrongyliasis.
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Adult , Humans , Angiostrongylus cantonensis , Angiostrongylus , Antibodies, Monoclonal , Diagnosis , Enzyme-Linked Immunosorbent Assay , Immunoassay , Limit of Detection , Methods , Sensitivity and Specificity , Serologic TestsABSTRACT
<p><b>OBJECTIVE</b>To develop a novel sensitive duplex real-time PCR assay for accurately identifying B/Yamagata and B/Victoria lineages of influenza virus type B.</p><p><b>METHODS</b>50 HA (hemagglutinin) gene sequences coding for B/Yamagata and B/Victoria lineage, respectively, were randomly downloaded for GenBank and analyzed by software MEGA. Primers and probes specific for HA gene of B/Yamagata and B/Victoria lineages were designed by Primer Primer and then applied in the duplex real-time RT-PCR method that was followed developed. Influenza virus B type and A type isolated in our laboratory and typing-confirmed by HAI method were used as reference strains to determine the specificity of this assay and the sensitivity of the duplex amplification was evaluated by viral load testing in terms of in vitro transcribed RNA copy number.</p><p><b>RESULTS</b>In 2006-2010, 793 influenza virus type B strains were isolated from 17 765 throat swab samples, among which 152 strains were differentiated as By lineage and 641 as Bv lineage by this assay. These results was agreement with that determined by HAI assay. This developed assay allows to accurately identify approximately 10(2) copies/microl for Bv and By lineage virus with intra- and inter-coefficient of variation (CV) < 3.5% and nearly 100% specificity.</p><p><b>CONCLUSIONS</b>This method provides sensitive and robust tool for routine diagnosis and on-time epidemiological examination of influenza virus, which could be applied in influenza surveillance laboratories for rapid molecular diagnosis.</p>
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Hemagglutinin Glycoproteins, Influenza Virus , Genetics , Influenza B virus , Genetics , Real-Time Polymerase Chain Reaction , Methods , Sensitivity and SpecificityABSTRACT
<p><b>OBJECTIVE</b>Real-time RT-PCR, cell culture and embryonated eggs culture for influenza detection were compared by analyzing the data of influenza surveillance in Shenzhen in second half of 2009.</p><p><b>METHODS</b>1092 clinical samples (throat swabs) collected during second half of 2009 were tested by real-time RT-PCR, cell culture and embryonated eggs culture, and the results were analyzed by statistical methods.</p><p><b>RESULTS</b>The positive rate were 54.21%, 27.11% and 16.21% using real-time RT-PCR, cell culture and embryonated eggs culture, and the sensitive were 100%, 50% and 29.9%. The lowest dilutions of virus detected by real-time RT-PCR were 10(-2) TCID50/ml.</p><p><b>CONCLUSION</b>The sensitive of real-time RT-PCR was higher than culture and the specificity was also very high. It was more suitable for emergency detect. The sensitive of cell culture for H3N2 subtype was higher, and sensitive of embryonated eggs culture for type B was higher.</p>
Subject(s)
Animals , Chick Embryo , Humans , Orthomyxoviridae , Reverse Transcriptase Polymerase Chain Reaction , Methods , Virus Cultivation , MethodsABSTRACT
Objective To detect the rate of Angiostrongylus cantonensis infection and to study the effects of treatment so as to prepare monoclonal antibodies (McAbs) ,and gold immunochroma-tography assay (GICA) with 12D5 and 21B7 McAbs could be prepared in advance. Methods Two McAbs (12D5 and 21B7) were applied to detect the circulating antigen (CAg) in the sera of rats infected with A. cantonensis and angiostrongyliasis patients respectively by double antibody sandwich ELISA. Either 12D5 or 21B7 McAbs was used as antibody and protein A was conjugated with colloid gold as the detection marker. A special pad for GICA was designed according to the reaction procedure, and CAg were detected by GICA in the sera of rats infected with A. cantonensis and angiostrongyliasis patients respectively. Results 12D5 McAb was identified as IgG1 and 21B7 McAb was IgM. Results from Western blotting showed that two McAbs could be used to identified 55 KD protein of adult worms of A. cantonensis. The detection rates of CAg in the sera of infected rats was 100% (48/48) and the detection rates of CAg in the sera of angiostrongyliasis patients was 100% (32/32). No cross-reaction to sera of patients with other infection of parasites, such as clonochiasis, fasiolopsiasis, ancylostotniasis, ancylostomiasis, anisakiasis as well as schsitosomiasis wee seen and normal sera did not react with 12D5 and 21B7 McAbs. Conclusion Results from sandwich ELISA and GICA with 12D5 and 21B7 McAbs showed high specificity and acting as detecting CAg of A. cantonensis in sera of infected animals and patients. We noticed that GICA with 12D5 and 21B7 was not only rapid and simple that without requirement of special instrument, but also rather sensitive and specific for the detection of current infection with A.cantonensis.
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Analysis of serological and genetic characteristics on 2009 swine-origin influenza A (H1N1) virus (S-OIV) isolated from four patients with severe disease in Shenzhen were performed. Microneutralization assay showed that the neutralizing antibody titers of the infected patients did not exceed 1 : 20 in a short term post infection, which could not neutralize the viruses efficiently. Hemagglutination inhibition (HI) tests confirmed that the antigenicity of S-OIV from the patients was distinct from the seasonal influenza A virus, but similar to the reference strains of S-OIV. Phylogenetic and molecular analysis showed that S-OIV from the patients still belonged to the classical swine lineages and did not have the genetic characteristics of highly pathogenic influenza virus. Several amino acid residue mutations on HA protein were detected, which seemed not to affect the virulence and pathogenicity of the viruses. Further, A His 275 Tyr mutation on NA protein of a virus strain was detected, which induced the oseltamivir resistance of the virus.
Subject(s)
Child , Child, Preschool , Female , Humans , Male , Young Adult , Amino Acid Sequence , Antibodies, Neutralizing , Allergy and Immunology , Antibodies, Viral , Allergy and Immunology , China , Influenza A Virus, H1N1 Subtype , Classification , Genetics , Influenza, Human , Allergy and Immunology , Virology , Molecular Sequence Data , Mutation , Phylogeny , Sequence Alignment , Viral Proteins , Chemistry , GeneticsABSTRACT
Objective To explore the association between polymorphisms of adiponectin gene and the risk of ischemic stroke in Han population from the Northern parts of China.Methods TaqMan probe of RT-PCR was applied to detect the genotype frequency of single nucleotide polymorphism(SNPs)(rs266729 and rs2241766)of adiponectin gene in 357 ischemic stroke cases who developed the episode at first time and with 345 healthy controls.Logistic regression analysis was used to evaluate the relationship of each genotype of SNPs and ischemic stroke.Results Mutation of rs2241766(T＞G)increased the risk of ischemic stroke among all the samples(0R=1.55,P=0.01)and it was still the risk factor of ischemic stroke when analyzed by multi-factors logistic regression after each factor was adjusted(OR=1.55,P=0.00).The polymorphism of rs266729 was not related to the risk of ischemic stroke among all the samples(OR=1.13,P=0.57).However,the genotype GG of rs266729 increased the risk of ischemic stroke among female population(OR=3.25,P=0.04).Conclusion The variance of rs2241766 in adiponectin gene was related to the risk of ischemic stroke in Han population from the Northern parts of China and the genotype GG of rs266729 could possibly increase the risk of ischemic stroke in women of Han population from the Northern parts of the country.